葡萄Trihelix转录因子家族生物信息及其基因表达分析

Bioinformatics Identification and Expression Analysis of Grape Trihelix Transcription Factor Family

利用生物信息学分析、预测葡萄Trihelix转录因子家族基因的特性与功能,为葡萄抗逆基因的挖掘和利用提供理论依据。以‘红地球’葡萄(Vitis vinifera)试管苗为试验材料,对葡萄Trihelix转录因子家族进行理化性质、染色体定位、二级结构预测、亚细胞定位、基因结构分析、motif以及基因芯片表达分析,并利用qRT-PCR技术分析该家族基因在400 mmol·L^-1 NaCl、100 mmol·L^-1 ABA和10%PEG逆境下的表达情况。结果表明,该转录因子家族在葡萄基因组中总共有27个成员,分布于11条染色体上,其中有10个成员集中分布在第14号和17号染色体上;氨基酸残基为175~880个,大多数具有疏水性。亚细胞定位分析表明,该基因家族主要存在于细胞核中。结构域分析显示,同一亚家族蛋白保守域结构高度相似。基因芯片表达结果显示,非生物胁迫下,叶片中VvTrihelix6在PEG处理24 h后与对照相比呈显著上调趋势,当用NaCl处理4h和24h后,与叶片对照相比有下调表达趋势;用ABA处理果实3d后,该基因在果实中呈下调表达,处理10d后变化不明显。叶片中VvTrihelix19转录因子经PEG、NaCl和冷胁迫处理24 h后明显下调;果实用ABA处理3 d和10 d后均有明显下调趋势。qRT-PCR分析表明该转录因子家族所有基因对逆境胁迫具有响应作用,但是,不同成员在不同逆境胁迫下的响应程度存在一定的差异。转录因子VvTrihelix6在400 mmol·L^-1 NaCl和100 mmol·L^-1 ABA处理下表达量极显著高于对照,分别是对照的15倍和8倍;VvTrihelix19只在10%PEG胁迫处理后呈显著上调表达。

The characteristics and functions of grape Trihelix transcription factor family genes were analyzed and predicted by bioinformatics,providing a theoretical basis for the mining and utilization of grape stress resistant gene. In this study,Vitis vinifera was used as the material. Bioinformatics was used to analyze the physicochemical properties,chromosomal localization,secondary structure prediction,subcellular localization,gene structure analysis,motif explores and gene chip expression of grape Trihelix transcription factor family. The qRT-PCR was used to analyze the expression of this family under 400 mmol · L^-1 NaCl,100 mmol · L^-1 ABA and 10% PEG stress. The results showed that there were 27 members of the transcription factor family in the grape genome,and distributed on 11 chromosomes,including 10 members concentrated on chromosomes 14 and 17. The amino acid size is mainly between 175–880 aa,most is hydrophobic. The subcellular localization analysis indicated that this gene family mainly existed in the nucleus. Domain analysis displayed that the conserved domain structure of the same subfamily was highly similar. Gene chip expression results clearly illuminated that VvTrihelix6 played up-regulated trend after 24 h PEG treatment compared with the control under abiotic stress,and a down-regulated trend after 4 h and 24 h PEG treatment contrast with the control. Also,after ABA treatment for 3 d,the VvTrihelix6 was down-regulated in fruits,but the change was not obvious after 10 d. The transcription factor VvTrihelix19 showed a down regulation trend after 24 h treatment with PEG,salt and cold compare with control,and showed significant down-regulation at 3 d and 10 d under ABA treatment. qRT-PCR showed that all genes in the Trihelix transcription factor family responded to stress. However,the response degree of members of the gene family was different under different stress conditions. The expression of transcription factor VvTrihelix6 in 400 mmol · L^-1 NaCl and 100 mmol · L^-1 ABA treatment was significantly higher than the control group,15 times and 8 times of the control,respectively. VvTrihelix19 was significantly up-regulated after 10% PEG stress treatment.